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1.
Int Arch Allergy Immunol ; 181(2): 128-135, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31805564

RESUMO

BACKGROUND: In contrast to the 3 major aeroallergens tree pollen, grass pollen, and house dust mites, allergic rhinitis caused by herbal pollen has received comparatively little attention in recent clinical studies. Since various weeds flower during summer until fall, allergic rhinitis to weeds may be underdiagnosed and/or mistakenly diagnosed as grass pollen allergy. OBJECTIVE: To investigate (i) the currently most frequent weed allergy between mugwort, ragweed, plantain, chamomile, nettle, and oilseed rape and (ii) time trends in prevalence of sensitization to weed pollen in the middle of Germany over the last 20 years. METHODS: This study, the largest of its kind to date, monocentrically evaluated the prick test results of a total of 6,220 patients with suspected RCA over a period of 20 years (1998-2017). RESULTS: In the study cohort, sensitization rates to plantain almost doubled from 26.6% in the decade 1998-2007 to 50.5% in 2008-2017. Identical increases were observed for ragweed, while sensitization rates for mugwort stayed largely unchanged. The most prominent increase in positive skin prick tests to plantain and ragweed pollen was mainly observed in younger patients. Further, we identified a trend toward polysensitization, currently dominated by plantain and ragweed. Sensitization to weed pollen was found to be highly associated with additional sensitizations to grass and/or birch pollen. CONCLUSION: Plantain is currently the best choice to screen rhinitis patients for weed allergy which identifies 86% of all weed-sensitized individuals, at least in Germany. Over the last 20 years, we demonstrate a significant rise in the total number of weed pollen sensitization as well as increases in polysensitization, predominantly in younger patients.


Assuntos
Alérgenos/imunologia , Ambrosia/imunologia , Plantago/imunologia , Pólen/imunologia , Rinite Alérgica/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos de Plantas/imunologia , Artemisia/imunologia , Criança , Pré-Escolar , Feminino , Alemanha , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/imunologia , Prevalência , Estudos Retrospectivos , Rinite Alérgica Sazonal/imunologia , Testes Cutâneos/métodos , Adulto Jovem
2.
Food Chem ; 307: 125565, 2020 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-31630022

RESUMO

The effectiveness of some non-specific proteases in reducing raw peanut allergenicity was investigated. Peanut kernels were treated by Alcalase, papain, Neutrase and bromelain, respectively. The residues of major peanut allergens Ara h 1, Ara h 2 and Ara h 6 were determined by sandwich ELISA and SDS-PAGE, and the allergenicities of treated peanuts were compared to that of untreated peanuts by western blot. All tested proteases were effective in reducing Ara h 1, but their effectiveness in hydrolyzing Ara h 2 and Ara h 6 varied greatly. The maximal reductions of extractable Ara h 1, Ara h 2 and Ara h 6 were 100%, 100% and 99.8%, respectively, achieved by Alcalase hydrolysis. Alcalase was more effective in overall allergenicity reduction; bromelain and Neutrase were the least effective in reducing Ara h 2 and Ara h 6, respectively. The hydrolysis of original allergens also produced some smaller peptides with strong IgE-binding.


Assuntos
Alérgenos/metabolismo , Arachis/química , Imunoglobulina E/imunologia , Hipersensibilidade a Amendoim/prevenção & controle , Peptídeo Hidrolases/metabolismo , Albuminas 2S de Plantas/análise , Albuminas 2S de Plantas/imunologia , Albuminas 2S de Plantas/metabolismo , Alérgenos/análise , Alérgenos/imunologia , Antígenos de Plantas/análise , Antígenos de Plantas/imunologia , Antígenos de Plantas/metabolismo , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Humanos , Proteínas de Membrana/análise , Proteínas de Membrana/imunologia , Proteínas de Membrana/metabolismo , Proteínas de Plantas/análise , Proteínas de Plantas/imunologia , Proteínas de Plantas/metabolismo
3.
Int Arch Allergy Immunol ; 181(2): 94-102, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31865326

RESUMO

Only few data on safety during high-dose, accelerated escalation schedules during subcutaneous allergen immunotherapy (AIT) are available. The aim of this study was to assess the safety and tolerability of an accelerated dose escalation schedule of AIT in adult patients with moderate to severe seasonal rhinoconjunctivitis in a multicenter, open-label, randomized phase II trial. The dose escalation scheme for patients in Group I (1 strength) included 3 injections with 1 strength, B (10,000 TU/mL), whereas the dose escalation scheme for Group II (standard) included 7 injections with 2 strengths, A (1,000 TU/mL) and B (10,000 TU/mL), of an aluminum hydroxide-adsorbed allergoid grass pollen preparation. Overall, 72 of 87 randomized patients (83.7%) reported at least 1 treatment-emergent adverse event (TEAE; 82.2 [Group I] vs. 85.4% [Group II]); 58.8% of all reported TEAEs were assessed as being related to AIT (60.0 vs. 48.8%). The most frequently reported AIT-related TEAEs were swelling (46.7 vs. 34.1%), erythema (28.9 vs. 36.6%), and pruritus (31.1 vs. 17.1%) at the site of the injection. Systemic allergic reactions occurred in 5 (5.8%) patients overall, with more being reported in the 1-strength group (4 [8.9%] vs. 1 [2.4%]). All systemic allergic reactions were classified as World Allergy Organization (WAO) Grade 1 or Grade 2 reactions. Accelerated high-dose escalation with an aluminum hydroxide-adsorbed grass pollen allergoid can be initiated with a safety and tolerability profile comparable to the standard dose escalation schedule in patients with allergic rhinitis with or without asthma.


Assuntos
/química , Hidróxido de Alumínio/química , Poaceae/imunologia , Rinite Alérgica Sazonal/imunologia , Rinite Alérgica/imunologia , Adulto , Alérgenos/imunologia , Hidróxido de Alumínio/administração & dosagem , Hidróxido de Alumínio/imunologia , Antígenos de Plantas/imunologia , Asma/imunologia , Conjuntivite Alérgica/imunologia , Dessensibilização Imunológica/métodos , Feminino , Humanos , Masculino , Pólen/imunologia
4.
J Agric Food Chem ; 67(47): 13127-13138, 2019 Nov 27.
Artigo em Inglês | MEDLINE | ID: mdl-31682438

RESUMO

Allergen-specific immunotherapy is the only available curative treatment for IgE-mediated allergen diseases. A safe hypoallergenic allergen derivative with high efficiency is required as a tolerogen to induce immune tolerance to the causitive allergens. In this study, to generate a rice-based oral allergy vaccine for Japanese cedar (JC) pollinosis, the tertiary structures of major JC pollen allergens, Cry j 1 and Cry j 2, were more completely destructed by shuffling than the previous ones without losing immunogenicity and then were specifically expressed in the endosperm of transgenic rice seed. They accumulated at high levels and were deposited in endoplasmic reticulum (ER) and ER-derived protein bodies. The low allergenicity of these deconstructed Cry j 1 and Cry j 2 allergens was evaluated by examining their binding activities to the specific IgE antibody and by the basophil degranulation test.


Assuntos
Antígenos de Plantas/imunologia , Cryptomeria/imunologia , Hipersensibilidade/imunologia , Oryza/genética , Plantas Geneticamente Modificadas/genética , Animais , Antígenos de Plantas/genética , Cryptomeria/genética , Humanos , Hipersensibilidade/terapia , Imunoterapia , Camundongos , Oryza/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Pólen/genética , Pólen/imunologia , Ratos , Sementes/genética , Sementes/imunologia , Vacinas/administração & dosagem , Vacinas/genética , Vacinas/imunologia
5.
Mol Immunol ; 116: 199-207, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31731097

RESUMO

A 38 kDa ß-1,3-glucanase allergen from Cryptomeria japonica pollen (CJP38) was recombinantly produced in E. coli and purified to homogeneity with the use of Ni-affinity resin. CJP38 hydrolyzed ß-1,3-glucans such as CM-curdlan and laminarioligosaccharides in an endo-splitting manner. The optimum pH and temperature for ß-1,3-glucanase activity were approximately 4.5 and 50 °C, respectively. The enzyme was stable at 30-60 °C and pH 4.0-10.5. Furthermore, CJP38 catalyzed a transglycosylation reaction to yield reaction products with a molecular weight higher than those of the starting laminarioligosaccharide substrates. The three-dimensional structure of CJP38 was determined using X-ray crystallography at 1.5 Å resolution. CJP38 exhibited the typical (ß/α)8 TIM-barrel motif, similar to allergenic ß-1,3-glucanases from banana (Mus a 5) and rubber tree latex (Hev b 2). Amino acid sequence alignment of these proteins indicated that the two-consensus IgE epitopes identified on the molecular surfaces of Mus a 5 and Hev b 2 were highly conserved in CJP38. Their conformations and surface locations were quite similar for these proteins. Sequence and structural conservation of these regions suggest that CJP38 is a candidate allergen responsible for the pollen-latex-fruit syndrome relating to Japanese cedar pollinosis.


Assuntos
Alérgenos/química , Antígenos de Plantas/química , Cryptomeria/química , Pólen/química , Alérgenos/imunologia , Sequência de Aminoácidos , Antígenos de Plantas/imunologia , Reações Cruzadas/imunologia , Cryptomeria/imunologia , Cristalografia por Raios X/métodos , Epitopos/química , Epitopos/imunologia , Escherichia coli/imunologia , Humanos , Concentração de Íons de Hidrogênio , Imunoglobulina E/química , Imunoglobulina E/imunologia , Látex/química , Látex/imunologia , Musa/química , Musa/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Temperatura Ambiente
6.
Nat Immunol ; 20(12): 1692-1699, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31745340

RESUMO

High-throughput 3' single-cell RNA-sequencing (scRNA-seq) allows cost-effective, detailed characterization of individual immune cells from tissues. Current techniques, however, are limited in their ability to elucidate essential immune cell features, including variable sequences of T cell antigen receptors (TCRs) that confer antigen specificity. Here, we present a strategy that enables simultaneous analysis of TCR sequences and corresponding full transcriptomes from 3'-barcoded scRNA-seq samples. This approach is compatible with common 3' scRNA-seq methods, and adaptable to processed samples post hoc. We applied the technique to identify transcriptional signatures associated with T cells sharing common TCRs from immunized mice and from patients with food allergy. We observed preferential phenotypes among subsets of expanded clonotypes, including type 2 helper CD4+ T cell (TH2) states associated with food allergy. These results demonstrate the utility of our method when studying diseases in which clonotype-driven responses are critical to understanding the underlying biology.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Hipersensibilidade a Amendoim/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Células Th2/imunologia , Albuminas 2S de Plantas/imunologia , Animais , Antígenos de Plantas/imunologia , Células Cultivadas , Regiões Determinantes de Complementaridade/genética , Feminino , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Imunização , Imunoglobulina E/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Proteínas E7 de Papillomavirus/imunologia , Análise de Célula Única , Especificidade do Receptor de Antígeno de Linfócitos T/genética , Transcriptoma
7.
J Agric Food Chem ; 67(45): 12547-12557, 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31613616

RESUMO

This study aims at understanding the changes in the structural properties of Gly m 4 soy allergen as a result of the influence of the temperature and pressure deviations. The primary emphasis was placed on analyzing the surface properties of suspected linear and conformational epitopes present in the Gly m 4 allergen. All three epitopes of Gly m 4 were studied, and the results showed that the molecule has significant structural changes in terms of solvent-accessible surface area (SASA) and radius of gyration, which showed that the increased pressures resulted in compaction. However, at lower temperatures and higher pressures (300 K and 6 kbar), swelling in the molecule was observed with a significant increase in the surface area. The study also tracked the changes in surface areas of individual residues that are part of the selected epitopes. Residues, such as D-27 and T-51, were found to have significant changes in their SASA as a result of temperature and pressure deviations.


Assuntos
Alérgenos/química , Antígenos de Plantas/química , Soja/imunologia , Alérgenos/imunologia , Antígenos de Plantas/imunologia , Mapeamento de Epitopos , Modelos Moleculares , Pressão , Soja/química , Temperatura Ambiente
9.
Int Arch Allergy Immunol ; 180(3): 212-220, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31533105

RESUMO

BACKGROUND: Although plant and fruit pollens are entomophilous and relevant in exposed workers, we have shown a high frequency of sensitisation and symptoms induction of peach tree pollen (PTP) and Prunus persica 9 (Pru p 9) in adults from areas of peach cultivars. METHODS: We studied the sensitisation and clinical relevance of PTP and Pru p 9 in a large group of children and adolescents aged 3-19 years. A detailed questionnaire plus skin prick testing to prevalent allergens, PTP, and Pru p 9 were carried out. The clinical relevance was established by nasal provocation test (NPT) and symptom score index. RESULTS: We evaluated 685 children (mean age 8.75 ± 3.3 years, median 9 years), 52% of them female. Sensitisation to PTP occurred in 20% of the cases following olive tree (33%) and Phleum pratense (26%). In a randomly selected subgroup of subjects sensitised to PTP, 30% were skin prick test-positive to Pru p 9. Most cases had rhinitis or rhinoconjunctivitis. NPT showed the relevance of PTP and Pru p 9 in the induction of symptoms. CONCLUSION: PTP and Pru p 9 are relevant in the induction of sensitisation and respiratory symptoms in children and adolescents. This allergen should be evaluated in children living in regions of peach tree cultivars.


Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Hipersensibilidade Alimentar/epidemiologia , Hipersensibilidade Alimentar/imunologia , Pólen/imunologia , Prunus persica/imunologia , Rinite Alérgica Sazonal/imunologia , Adolescente , Criança , Feminino , Humanos , Imunização , Masculino , Olea/imunologia , Phleum/imunologia , Espanha/epidemiologia
10.
Mol Immunol ; 114: 189-195, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31376732

RESUMO

The presence in cypress pollen of an important allergen, belonging to the gibberellin-regulated protein (GRP) family, has been suggested for many years. However, it has never been isolated and sometimes the homologous peach allergen, Pru p 7, has been used as a surrogate to perform immunological investigations. The aim of this study has been the isolation and molecular characterization of the GRP contained in the Cupressus sempervirens pollen. This protein, named Cypmaclein, has been purified from the natural source using conventional biochemical methods consisting in different chromatographic separations. Cypmaclein has been identified by direct protein sequencing of the N-terminal region and of internal fragments of the molecule. In SDS-PAGE, its apparent molecular mass is slightly higher than that of Pru p 7. Nevertheless, the mass spectrometry experiments reveal that the exact molecular mass of Cypmaclein (6821.88 Da) is very close to that of Pru p 7 (6909.90 Da). Two regions of Cypmaclein have been sequenced providing 50% of its primary structure. A high overall sequence identity of Cypmaclein with all the analyzed GRP has been observed, although in the N-terminal region the high identity is limited to the homolog of Cryptomeria japonica. In circular dichroism experiments Cypmaclein produced a spectrum overlapping that of Pru p 7. However, the comparative analysis of Cypmaclein, Pru p 7 and Pun g 7 IgE reactivity revealed a behavior that was not completely overlapping, thus suggesting that the IgE epitopes are only partially shared. In single point highest inhibition achievable assays performed with the FABER test, Cypmaclein efficiently competed with the allergenic peach and pomegranate GRP in the binding of specific IgE of patients sensitized to Pru p 7. In conclusion, the natural cypress pollen GRP has been isolated for the first time, its structural features have been investigated and its cross-reactivity with Pru p 7 and Pun g 7 has been demonstrated. This protein is now available for further investigations aimed at understanding its clinical relevance in the allergy to cypress pollen. In addition, the prevalence of sensitization directly to Cypmaclein, and not limited to the homologs, can be defined.


Assuntos
Cupressus/química , Cupressus/imunologia , Giberelinas/química , Giberelinas/imunologia , Imunoglobulina E/imunologia , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Adolescente , Adulto , Sequência de Aminoácidos , Antígenos de Plantas/química , Antígenos de Plantas/imunologia , Criança , Reações Cruzadas/imunologia , Epitopos/química , Epitopos/imunologia , Feminino , Humanos , Masculino , Pólen/química , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Adulto Jovem
11.
Arerugi ; 68(7): 851-856, 2019.
Artigo em Japonês | MEDLINE | ID: mdl-31406081

RESUMO

BACKGROUND: The present study aimed to assess the safety of a three-level stepwise oral food challenge (OFC) in patients with peanut allergy. METHODS: This retrospective analysis reviewed laboratory test results and food allergy histories (obtained from medical records) of participants who underwent OFC using peanut butter for the first time at Miyagi Children's Hospital between January 2015 and December 2017. Total food dose was classified as follows: step 1, 0.1 g peanut butter; step 2, 0.5g peanut butter; and step 3, 3g peanut butter. RESULTS: A total of 114 participants were included. OFC-positive rates were 52% (step 1), 35% (step 2), and 20% (step 3). Of the 41 OFC-positive participants, 1 required an intramuscular adrenaline injection during step 3. Peanut allergy history rates and peanut- and Ara h 2-specific immunoglobulin E (sIgE) titers were significantly higher during the low-dose step. CONCLUSION: The three-level stepwise OFC with peanut butter is safe for patients with peanut allergy, as indicated by peanut- and Ara h 2-sIgE titers.


Assuntos
Alimentos , Hipersensibilidade a Amendoim/diagnóstico , Albuminas 2S de Plantas/imunologia , Antígenos de Plantas/imunologia , Arachis , Criança , Glicoproteínas/imunologia , Humanos , Imunoglobulina E/sangue , Estudos Retrospectivos
12.
J Food Sci ; 84(8): 2357-2363, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31364176

RESUMO

Peanut allergic consumers rely on food package labels to avoid foods containing peanut. The inadvertent presence of peanut in foods due to cross-contact can be fatal if ingestion of such food leads to an allergic reaction. Analytical methods are available to detect undeclared peanut in foods. However, depending on the type of food matrix and food processing parameters, method performance can be adversely affected due to reduction in the extraction efficiency of peanut proteins. Temperature and probe sonication were used as a preincubation treatment for peanut flour slurries to assess their effect on the total peanut protein solubility from raw, light-roasted, and dark-roasted peanut flours. The effect of these treatments on the immunoreactivity of peanut allergens (Ara h 1, Ara h 2, Ara h 3, and Ara h 6) was determined by an indirect enzyme-linked immunosorbent assay using antibodies raised against these individual peanut proteins. Preincubation at 50 °C did not significantly improve the peanut protein solubility, whereas an increase in protein solubility was observed when light- and dark-roasted peanut flour slurries were preincubated at 90 °C or sonicated. The immunoreactivity of peanut allergens varied depending on the degree of peanut flour roasting and type of preincubation treatment. Overall, the immunoreactivity of peanut allergens from most peanut flour slurries was unaffected when preincubated at 50 °C for up to 60 min or sonicated with a probe for up to 5 min, whereas preincubation at 90 °C resulted in a time-dependent reduction in immunoreactivity of peanut allergens. Sonication treatment may improve peanut protein extraction without markedly affecting their immunoreactivity. PRACTICAL APPLICATION: Extraction of peanut proteins is vital for developed analytical methods to estimate peanut allergens in foods. The manuscript describes the effect of two different temperatures (50 and 90 °C) and probe-type sonication on peanut protein solubility. The findings suggest sonication can improve peanut protein solubility without markedly affecting their immunoreactivity.


Assuntos
Arachis/imunologia , Antígenos de Plantas/análise , Antígenos de Plantas/imunologia , Antígenos de Plantas/isolamento & purificação , Arachis/química , Ensaio de Imunoadsorção Enzimática , Farinha/análise , Manipulação de Alimentos , Humanos , Proteínas de Plantas/análise , Proteínas de Plantas/imunologia , Proteínas de Plantas/isolamento & purificação , Temperatura Ambiente
13.
J Agric Food Chem ; 67(40): 11219-11229, 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31408330

RESUMO

Peanut allergy is a major health problem worldwide. Detection of food allergens is a critical aspect of food safety. The VHH domain of single chain antibody from camelids, also known as nanobody (Nb), showed its advantages in the development of biosensors because of its high stability, small molecular size, and ease of production. However, no nanobody specific to peanut allergens has been developed. In this study, we constructed a library with random triplets (NNK) in its CDR regions of a camel nanobody backbone. We screened the library with peanut allergy Ara h 3 and obtained several candidate nanobodies. One of the promising nanobodies, Nb16 was further biochemical characterization by gel filtration, isothermal titration calorimetry (ITC), cocrystallization, and Western blot in terms of its interaction with Ara h 3. Nb16 specifically binds to peanut major allergen Ara h 3 with a dissociation constant of 400 nM. Furthermore, we obtained the Ara h 3-Nb16 complex crystals. Structure analysis shows the packing mode is completely different between the Ara h 3-Nb16 complex crystal and the native Ara h 3 crystal. Structural determination of Ara h 3-Nb16 will provide the necessary information to understand the allergenicity of this important peanut allergen. The nanobody Nb16 may have application in the development of biosensors for peanut allergen detection.


Assuntos
Antígenos de Plantas/imunologia , Arachis/imunologia , Proteínas de Armazenamento de Sementes/imunologia , Anticorpos de Domínio Único/genética , Anticorpos de Domínio Único/imunologia , Sequência de Aminoácidos , Antígenos de Plantas/química , Antígenos de Plantas/genética , Arachis/química , Arachis/genética , Bacteriófagos/genética , Bacteriófagos/metabolismo , Western Blotting , Técnicas de Visualização da Superfície Celular , Proteínas de Armazenamento de Sementes/química , Proteínas de Armazenamento de Sementes/genética , Anticorpos de Domínio Único/análise
14.
Allergol. immunopatol ; 47(4): 357-364, jul.-ago. 2019. tab, graf
Artigo em Inglês | IBECS | ID: ibc-186507

RESUMO

Introduction: Amaranthus retroflexus (Redroot Pigweed) is one of the main sources of allergenic pollens in temperate areas. Polcalcin is a well-known panallergen involved in cross-reactivity between different plants. The aim of this study was the molecular cloning and expression of polcalcin, as well as evaluating its IgE-reactivity with A. retroflexus sensitive patients' sera. Methods: Allergenic extract was prepared from A. retroflexus pollen and the IgE-reactivity profile was determined by ELISA and immunoblotting using sera from twenty A. retroflexus sensitive patients. Polcalcin-coding sequence was amplified by conventional PCR method and the product was inserted into pET-21b(+) vector. The recombinant protein was expressed in E. coli BL21 and purified by metal affinity chromatography. The IgE-binding capability of the recombinant protein was analyzed by ELISA and immunoblotting assays, and compared with crude extract. Results: Of 20 skin prick test positive patients, 17 patients were positive in IgE-specific ELISA. Western blotting confirmed that approximately 53% of ELISA positive patients reacted with 10kDa protein in crude extract. The A. retroflexus polcalcin gene, encoding to 80 amino acid residues was cloned and expressed as a soluble protein and designated as Ama r 3. The recombinant polcalcin showed rather identical IgE-reactivity in ELISA and western blotting with 10 kDa protein in crude extract. These results were confirmed by inhibition methods, too. Conclusion: The recombinant form of A. retroflexus polcalcin (Ama r 3) could be easily produced in E. coli in a soluble form and shows rather similar IgE-reactivity with its natural counterpart


No disponible


Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto Jovem , Adulto , Alérgenos/imunologia , Amaranthus/imunologia , Antígenos de Plantas/imunologia , Proteínas de Ligação ao Cálcio/imunologia , Pólen/imunologia , Rinite Alérgica Sazonal/imunologia , Testes Cutâneos , Alérgenos/isolamento & purificação , Antígenos de Plantas/isolamento & purificação , Proteínas de Ligação ao Cálcio/isolamento & purificação , Clonagem Molecular , Reações Cruzadas , Escherichia coli/genética , Expressão Gênica , Imunoglobulina E/metabolismo , Extratos Vegetais , Proteínas Recombinantes/isolamento & purificação
15.
J Agric Food Chem ; 67(31): 8626-8631, 2019 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-31287307

RESUMO

An almond allergen with two known short peptide sequences was reported as the almond 2S albumin but was later suspected to be almond vicilin. However, this allergen was not designated by the World Health Organization/International Union of Immunological Societies. This study aimed to determine the true identity of this elusive almond allergen. cDNAs were synthesized from total RNA of the Nonpareil almond. The complete sequence of the previously reported almond allergen was determined from its coding sequence. The deduced protein was produced recombinantly and was confirmed to be a food allergen by testing with 18 almond-allergic sera. The allergen is a potential cysteine-rich antimicrobial protein with characteristic C[X]3C-[X]10-12-C[X]3C motifs of the hairpinin antimicrobial protein. This first member of a novel family of food allergens was named Pru du 8. The signature motif of the hairpinin antimicrobial protein can be found in the N-terminal region of some vicilin allergens (e.g., Ara h 1). It can also be found in the signal peptide of other vicilin allergens (e.g., Car i 2). In many species, however, vicilins do not contain such a motif, indicating that the presence of the signature motifs of the hairpinin antimicrobial protein in vicilins might be a result of translocation during evolution.


Assuntos
Alérgenos/imunologia , Antígenos de Plantas/imunologia , Prunus dulcis/imunologia , Alérgenos/química , Alérgenos/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Antígenos de Plantas/química , Antígenos de Plantas/genética , DNA Complementar/genética , Hipersensibilidade Alimentar/imunologia , Humanos , Prunus dulcis/química , Prunus dulcis/genética , Proteínas de Armazenamento de Sementes/química , Proteínas de Armazenamento de Sementes/genética , Proteínas de Armazenamento de Sementes/imunologia , Alinhamento de Sequência , Análise de Sequência de DNA
16.
J Agric Food Chem ; 67(32): 9009-9021, 2019 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-31319030

RESUMO

Soybean allergy is a serious health risk to humans and animals; ß-conglycinin is the primary antigenic protein in soybean. Intestinal porcine epithelial (IPEC-J2) cells were used as an in vitro physiological model of the intestinal epithelium to study the effects of different concentrations of soybean antigen protein ß-conglycinin to identify the involved signaling pathways. The cells were divided into eight groups and either untreated or treated with different concentrations of ß-conglycinin, pyrrolidine dithiocarbamate (PDTC), Nω-nitro-l-arginine methyl ester hydrochloride (l-NAME), SP600125, and SB202190 either alone or in combination. The cells were incubated with 1, 5, and 10 mg·mL-1 ß-conglycinin or 5 mg·mL-1 ß-conglycinin and 1 µmol·L-1 nuclear factor κB (NF-κB) inhibitor (PDTC), inducible nitric oxide synthase inhibitor (l-NAME), c-Jun N-terminal kinase (JNK) inhibitor (SP600125), and p38 inhibitor (SB202190) for 24 h, separately; controls were left untreated. The mRNA, protein, and phosphorylation levels of NF-κB, p38, and JNK were higher in the treated groups than in the control group. ß-Conglycinin decreased tight junction distribution, destroyed the cytoskeleton of IPEC-J2 cells, and caused cell death. After the addition of the inhibitors, ß-conglycinin-induced IPEC-J2 cell damage was significantly reduced. ß-Conglycinin caused damage to IPEC-J2 cells via the mitogen-activated protein kinase/NF-κB signaling pathway. The results of this study are crucial for exploring the mechanisms underlying allergic reactions caused by soybean antigen proteins.


Assuntos
Antígenos de Plantas/imunologia , Células Epiteliais/imunologia , Hipersensibilidade Alimentar/imunologia , Globulinas/imunologia , Proteínas Quinases Ativadas por Mitógeno/imunologia , NF-kappa B/imunologia , Proteínas de Armazenamento de Sementes/imunologia , Proteínas de Soja/imunologia , Soja/imunologia , Animais , Humanos , Mucosa Intestinal/citologia , Mucosa Intestinal/imunologia , Proteínas Quinases JNK Ativadas por Mitógeno/genética , Proteínas Quinases JNK Ativadas por Mitógeno/imunologia , Proteínas Quinases Ativadas por Mitógeno/genética , NF-kappa B/genética , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/imunologia , Fosforilação , Transdução de Sinais , Suínos , Junções Íntimas/genética , Junções Íntimas/imunologia
17.
Mol Immunol ; 114: 19-29, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31326654

RESUMO

Worldwide, more than one-third of the population suffers from allergies. A significant fraction of officially registered allergens originate from the profilin family of proteins. Profilins are small ubiquitous proteins which are found in plants, viruses and various eukaryotes including mammals. Although they are primarily regarded as minor allergens, profilins are important players in immunoglobulin E (IgE) cross-reactivity. However, in some populations profilins are recognized by IgE from at least 50% of patients allergic to a given allergen source. Cuc m 2.0101 is recognized by IgE in more than 80% of muskmelon-allergic patients. The recombinant isoallergen Cuc m 2.0101 was produced in significant quantities and its X-ray crystal structure was determined. In addition, a new Art v 4.0101 (mugwort profilin) structure was determined. The profilins Cuc m 2.0101 and Art v 4.0101 were compared in terms of their structure and thermal stability. Furthermore, structural similarities and IgE cross-reactivity between profilins from different sources are discussed to explain the molecular basis of various clinical syndromes involving this group of allergens. Special emphasis is placed on discussion of profilins' quaternary structures and their relation to biological function, as well as to protein allergenicity. Moreover, a potential impact of protein purification protocols on the structure of profilins is highlighted.


Assuntos
Antígenos de Plantas/química , Profilinas/química , Sequência de Aminoácidos , Antígenos de Plantas/imunologia , Reações Cruzadas/imunologia , Escherichia coli/imunologia , Escherichia coli/metabolismo , Hipersensibilidade/imunologia , Imunoglobulina E/química , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Pólen/química , Pólen/imunologia , Profilinas/imunologia , Proteínas Recombinantes/química , Proteínas Recombinantes/imunologia
18.
Int J Biol Macromol ; 138: 97-105, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31302128

RESUMO

The inhalation of Chenopodium album (C. album) pollen, especially polcalcin (Che a 3), has been reported as a significant source of allergic respiratory symptoms. This study was conducted to characterize biophysical differences of recombinant polcalcin come from three different Escherichia coli strains using attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), optimize recombinant polcalcin expression, and linear B-cell epitopes identification using in silico methods. ATR-FTIR results of purified proteins showed spectra intensity variance in the amide I region, while there were no changes in pick position and shape of the bands. SHuffle® T7 Express was selected for subsequent optimization due to ability in the correction of the mis-oxidized bonds and promotes proper folding which was validated by ATR-FTIR analysis results. Then, Response Surface Methodology was performed to optimize critical factors including induction temperature, duration of induction, and concentration of inducer. The best partitioning conditions were achieved in 1.5 mM IPTG for 10.97 h at 29.9 °C. Finally, prediction of polcalcin B-cell epitopes was carried out which indicated the presence of 4 different epitopes. Together, the results may help to the development of diagnostic approaches and also vaccine manufacture for desensitization and modulation of the allergic response in patients.


Assuntos
Antígenos de Plantas/genética , Antígenos de Plantas/imunologia , Proteínas de Ligação ao Cálcio/genética , Proteínas de Ligação ao Cálcio/imunologia , Simulação por Computador , Escherichia coli/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Espectroscopia de Infravermelho com Transformada de Fourier , Antígenos de Plantas/química , Sítios de Ligação , Proteínas de Ligação ao Cálcio/química , Mapeamento de Epitopos , Expressão Gênica , Imunoglobulina E/imunologia , Peso Molecular , Proteínas Recombinantes/química
20.
Immunology ; 158(2): 94-103, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31323138

RESUMO

Transgenic rice seeds that contain genetically modified Cry j 1 and Cry j 2, the two major allergens of Cryptomeria japonica (Japanese cedar; JC), have been developed as immunotherapeutic candidates for JC pollinosis. Because the transgenic rice (TG-rice) seeds express allergens containing whole amino acid sequences of Cry j 1 and Cry j 2 in the endosperm tissue (edible part of rice grain), they can potentially target all Cry j 1- and Cry j 2-specific T-cells. However, it was unknown whether antigenicity of Cry j 1 and Cry j 2 could be completely preserved in TG-rice seeds. We verified the antigenicity of TG-rice seeds to T-cells through the analysis of the proliferative responses of T-cells in Cry j 1- or Cry j 2-immunized mice or T-cell lines to TG-rice seed extract. First, four mouse strains were immunized with Cry j 1 or Cry j 2. T-cells in the immunized mice proliferated on treatment with TG-rice seed extract, but not non-transgenic wild-type rice (WT-rice) seed extract. Furthermore, T-cell lines were established from the spleen cells of the immunized mice. Each T-cell line resulted in a proliferative response to TG-rice seed extract, but not to WT-rice seed extract, suggesting that TG-rice seeds certainly express T-cell epitopes corresponding to T-cell lines. Considering the modified amino acid sequences of Cry j 1 and Cry j 2 in TG-rice seeds, the expression of specific T-cell epitopes suggested that TG-rice seeds express all possible T-cell epitope repertoires of Cry j 1 and Cry j 2.


Assuntos
Alérgenos/farmacologia , Antígenos de Plantas/imunologia , Epitopos de Linfócito T/imunologia , Oryza/química , Proteínas de Plantas/imunologia , Rinite Alérgica Sazonal/imunologia , Linfócitos T/efeitos dos fármacos , Alérgenos/genética , Alérgenos/imunologia , Sequência de Aminoácidos , Animais , Antígenos de Plantas/química , Antígenos de Plantas/genética , Proliferação de Células/efeitos dos fármacos , Cryptomeria/genética , Cryptomeria/imunologia , Modelos Animais de Doenças , Epitopos de Linfócito T/química , Epitopos de Linfócito T/genética , Expressão Gênica , Imunização , Ativação Linfocitária/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos , Oryza/genética , Oryza/imunologia , Mapeamento de Peptídeos , Extratos Vegetais/imunologia , Extratos Vegetais/farmacologia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Pólen/genética , Pólen/imunologia , Cultura Primária de Células , Rinite Alérgica Sazonal/induzido quimicamente , Rinite Alérgica Sazonal/genética , Rinite Alérgica Sazonal/patologia , Sementes/química , Baço/efeitos dos fármacos , Baço/imunologia , Baço/patologia , Linfócitos T/imunologia , Linfócitos T/patologia , Transgenes
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